# Expression of Prooncogenic Nuclear Receptor 4A (NR4A)-Regulated Genes β1-Integrin and G9a Inhibited by Dual NR4A1/2 Ligands

## TL;DR

This study shows that dual NR4A1/2 ligands inhibit the expression of β1-integrin and G9a genes, which are regulated by NR4A1 and NR4A2 in colon cancer cells.

## Contribution

The study identifies dual NR4A1/2 ligands as inhibitors of NR4A-regulated genes and reveals shared and unique gene regulation by these receptors.

## Key findings

- DIM-3,5 compounds inhibit NR4A1- and NR4A2-regulated gene expression by displacing nuclear factors from gene promoters.
- NR4A1 and NR4A2 regulate both common and unique genes in SW480 colon cancer cells.
- Functional analysis shows that differentially expressed genes converge on shared pathways and gene ontology terms.

## Abstract

Bis-indole-derived compounds including 1,1-bis(3′-indolyl)-1-(3,5-disubstitutedphenyl)methane (DIM-3,5) analogs bind both orphan nuclear receptors 4A1 (NR4A1) and NR4A2, and DIM-3,5 compounds act as dual receptor inverse agonists and inhibit both NR4A1- and NR4A2-regulated responses. Chromatin immunoprecipitation assays show that β1-integrin and the methyltransferase gene G9a are regulated by both NR4A1 and NR4A2 acting as cofactors for Sp1- and Sp4-dependent gene expression. DIM-3,5 treatment results in the loss of one or more of these nuclear factors from the β1-integrin and G9a promoters. Single-cell and RNAseq analyses show that both receptors regulate common (<10%) and unique genes in SW480 colon cancer cells; however, functional enrichment analysis of the differentially expressed genes converges to several common pathways and gene ontology terms.